南京林业大学学报(自然科学版) ›› 1999, Vol. 23 ›› Issue (04): 7-12.doi: 10.3969/j.jssn.1000-2006.1999.04.002

• 研究论文 • 上一篇    下一篇

林木RAPD分析及实验条件的优化

尹佟明;韩正敏;黄敏仁;王明庥;李淑娴   

  1. 南京林业大学林木遗传与基因工程国家林业局重点实验室;南京林业大学森林资源与环境学院
  • 出版日期:1999-08-18 发布日期:1999-08-18

RAPD Experiment Protocol and Optimization in Forest Trees

Author Yin TongmingHan ZhengminHuang MinrenWang MingxiuLi Shuxian   

  1. Forest Genetics and Gene Engineering Laboratory Nanjing Forestry UniversityNanjing210037
  • Online:1999-08-18 Published:1999-08-18

摘要: <正>随机扩增多态DNA(RAPD,RandomAmplifiedPolymorphicDNA)分子标记是目前林木遗传研究中使用最为广泛的一种分子标记,作者对影响RAPD实验的因素进行了分析和探讨,确定了一个针对林木进行RAPD分析的实验程序。根据该程序,可以快速确定RAPD实验的理想条件,获得RAPD反应的良好重复性。

Abstract: RAPD marker is the most widely adopted molecular marker in forest genetic studies. The main reason is that this kind of markers can be obtained without any DNA sequence information about the studying material before hand. So it is especially suitable for the organism which little molecular study has been carried on. We can design primers arbitratily according to a common rule and the experiment process is relatively simple compared with other kinds of markers, such as RFLP and SSR. But this kind of marker has a fatal shortcoming, if the reaction condition is not suitable for the studying material, it gives out very flexible bands and cant be repeated. So the suitable amplification condition is critical to RAPD analysis. In this paper, the influencing factor on RAPD experiment was analyzed, such as the annealing temperature, the Mg2+ 2]concentration, the template concentration and so on. An optimized protocol especially suitable for forest trees was put forward through a series of experiments. Accord

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